[摘 要] 目的：探讨DNA甲基转移酶1（DNMT1）通过稳定zeste基因增强子同源物2（EZH2）促进结直肠癌（CRC）HCT8细胞增殖与迁移的机制。方法：利用生物信息学方法分析DNMT1在CRC组织中的表达水平。WB法检测DNMT1在CRC细胞HCT8、SW620和正常结肠上皮细胞NCM460中的表达。通过siRNA或慢病毒载体转染HCT8细胞，分为siNC组、siDNMT1组、Vector组、DNMT1-OE组、siTRAF6组、siEZH2组、siEZH2 + DNMT1-OE组。采用克隆形成实验、CCK-8法、Transwell实验和划痕愈合实验检测敲低或过表达DNMT1对HCT8细胞增殖与迁移的影响，WB和qPCR法检测EZH2蛋白和mRNA水平，免疫沉淀（IP）法检测EZH2泛素化水平，免疫荧光双染检测肿瘤坏死因子受体相关因子6（TRAF6）与EZH2的细胞内共定位情况，克隆形成和划痕愈合实验验证EZH2对DNMT1功能的逆转作用。收集2022—2025年间郑州大学附属洛阳中心医院手术切除的12例CRC患者的癌及癌旁组织标本，采用免疫组化法检测CRC组织中DNMT1、TRAF6和EZH2的表达水平。结果：DNMT1在CRC组织中表达显著高于癌旁组织（P < 0.01），且在CRC细胞中表达上调（P < 0.05）；DNMT1敲低显著抑制HCT8细胞增殖及迁移（均P < 0.01），过表达则相反（均P < 0.01）。DNMT1正向调控EZH2的蛋白水平（P < 0.01），而mRNA水平不变（P > 0.05）。MG132可恢复siDNMT1组的EZH2蛋白表达（P < 0.01），且siDNMT1组EZH2泛素化水平升高。DNMT1负向调控TRAF6的表达（P < 0.01），且TRAF6与EZH2在细胞质中共定位，IP证实两者直接结合。敲低TRAF6可减弱EZH2的泛素化水平，敲低EZH2可逆转DNMT1对HCT8细胞增殖、迁移的促进作用（均P < 0.01）。DNMT1和EZH2在CRC组织中呈高表达（P < 0.01），TRAF6在CRC组织中表达显著低于癌旁组织（P < 0.05）。结论：DNMT1通过抑制TRAF6稳定EZH2促进CRC细胞的增殖和迁移，DNMT1、TRAF6和EZH2可能是CRC治疗的潜在靶点。

ObjectiveTo establish a model of dry eye with lung Yin deficiency syndrome in mice. MethodsA total of 40 SPF C57BL/6J mice were assigned via the random number table method into 5 groups （n=8）： Normal control， model control， and high-， medium-， and low-dose （11.7， 5.85， and 2.925 g·kg^-1， respectively） Yangyin Qingfeitang. Mice in the normal control group were fed normally without any intervention. Mice in Yangyin Qingfeitang group and model control group were treated with 0.2% benzalkonium chloride eye drops （5 μL） twice a day and fed in a controlled drying system in a dry environment for 28 days. At the same time， the mice were administrated with thyroxine tablet solution by gavage and placed in a glass fumigation tank （SO₂ concentration： 0.5 g·m^-3） for 14 days. After 4 weeks， mice in Yangyin Qingfeitang groups were treated with Yangyin Qingfeitang by gavage and those in the normal control group and model control group were administrated with deionized water at 0.01 mL·g^-1. The body mass， anal temperature， four examination information （claw and nail appearance）， basic tear secretion test， tear film rupture time， corneal fluorescein staining， and lacrimal gland HE staining were compared among groups. Compound Yangyin Qingfeitang granules were used to measure the syndrome to verify the success of modeling. ResultsAfter 28 days of continuous modeling， compared with the normal control group， the model group exhibited listless and emaciated status， coughing， drowsiness， dry and dull hair， dry and hard stool， reduced food intake and water intake， red lip circumference， red tongue with reduced fluid， dry nose and teeth， red claws and nails， body mass gain， decreased anal mild tear secretion （P<0.05）， and shortened tear film rupture time （P<0.05）. After 28 days of modeling， the mice showed large corneal fluorescein staining range， severe corneal injury， and increased content of interleukin （IL）-18， IL-β， and tumor necrosis factor （TNF）-α in lacrimal gland， compared with those in the normal control group （P<0.05）. After the treatment with Yangyin Qingfeitang， the mice had good drinking and eating conditions， with lighter redness of the tongue， moist nose， moist and shiny teeth， and the claw and nail color close to that in the normal group. Compared with the model control group， Yangyin Qingfeitang groups showed increases in body mass and anal temperature （P<0.05）， tear secretion （P<0.05）， and tear film rupture time （P<0.05）， narrowed range of corneal fluorescein staining， and declined levels of IL-18， IL-β， and TNF-α in lacrimal glands （P<0.01）. The high-dose group had the best effect， with the indicators close to the levels in the normal control group. ConclusionThe animal model of dry eye with lung Yin deficiency syndrome can be established by culture in a controlled drying system， treatment with benzalkonium chloride eye drops for 28 days， and administration of thyroxine tablet solution combined with SO₂ fumigation for 14 days.

ObjectiveTo investigate the mechanisms of Runmu Dihuang decoction （RMDHD） in treating perimenopausal dry eye with liver-kidney Yin deficiency syndrome based on the silent information regulator 3 （SIRT3）/hypoxia-inducible factor-1α （HIF-1α）/nuclear factor-κB （NF-κB） signaling pathway. MethodsSixty female Sprague-Dawley rats were randomly divided into six groups （n=10 per group）： Sham operation group， model group， sodium hyaluronate eye drop group， and low-， medium-， and high-dose RMDHD groups （5.625， 11.25， 22.50 g·kg^-1）. Except for the sham operation group， all rats underwent bilateral ovariectomy and were administered 0.1% benzalkonium chloride eye drops combined with long-term chronic irritation to establish a perimenopausal dry eye model with liver-kidney Yin deficiency syndrome. Drug administration began in the 11th week after modeling and continued for 21 days. General conditions， screen-grip test scores， tear secretion volume， tear film breakup time （TFBUT）， and corneal fluorescein staining were recorded. Serum levels of reactive oxygen species （ROS）， follicle-stimulating hormone （FSH）， estradiol （E₂）， and progesterone （PROG） were measured by enzyme-linked immunosorbent assay （ELISA）. Pathological changes in the lacrimal glands， corneas， and uteri were observed using hematoxylin-eosin （HE） staining. Protein expression levels of SIRT3， HIF-1α， phosphorylated NF-κB p65 （p-NF-κB p65）， and total NF-κB p65 in the lacrimal glands were detected by Western blot. The expression of inflammatory cytokines interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） in the lacrimal glands was assessed by immunohistochemistry （IHC）. ResultsAfter model establishment， no significant differences were observed among the groups except the sham operation group. Compared with the sham operation group， the other groups exhibited slowed movement， dull responses， increased irritability， reduced body weight， elevated rectal temperature， decreased screen-grip test scores， reduced tear secretion， and significantly shortened TFBUT （P<0.05）. After treatment， compared with the model group， the sodium hyaluronate eye drop group and all RMDHD groups showed improved general conditions， significantly increased tear secretion （P<0.05）， prolonged TFBUT （P<0.05）， and elevated screen-grip test scores （P<0.05）. Serum ROS and FSH levels were significantly decreased， while E₂ and PROG levels were significantly increased （P<0.05）. Pathological damage to the cornea， lacrimal glands， and uterus was ameliorated. In addition， protein expression levels of SIRT3 and HIF-1α in the lacrimal glands were significantly upregulated （P<0.05）， whereas the expression of p-NF-κB p65， IL-1β， and TNF-α was significantly downregulated （P<0.05）. ConclusionRMDHD increases tear secretion and TFBUT， improves lacrimal gland and corneal injury， and alleviates dry eye symptoms in a perimenopausal dry eye rat model with liver-kidney Yin deficiency syndrome. The underlying mechanism may be related to regulation of the SIRT3/HIF-1α/NF-κB signaling pathway， inhibition of oxidative stress and inflammatory responses， and reduction of ocular surface tissue damage.

BackgroundChronic insomnia disorder has become a significant public health issue， and it may be associated with dyslipidemia. Previous studies on dyslipidemia in patients with chronic insomnia disorder have mainly focused on exploring the relationship between subjective short sleep duration and dyslipidemia， while there have been limited studies on the relationship between objective short sleep duration and dyslipidemia. ObjectiveTo explore the relationship between objective short sleep duration and dyslipidemia in patients with chronic insomnia disorder， in order to provide references for the prevention and intervention of dyslipidemia in this population. MethodsA total of 103 patients who were hospitalized at The Third Hospital of Mianyang from August 2022 to November 2023 and met the diagnostic criteria for chronic insomnia disorder as defined in the International Classification of Sleep Disorder， third edition （ICSD-3） were retrospectively collected. The objective sleep duration of the patients was obtained through polysomnography. The patients were divided into two groups based on their objective sleep duration： the group with objective sleep duration ≥ 7 hours （n=71） and the group with objective sleep duration < 7 hours （n=32）. Binary Logistic regression analysis was used to explore the impact of objective sleep duration < 7 hours on dyslipidemia. ResultsAmong 103 patients with chronic insomnia disorder， 59 cases （57.28%） were identified with dyslipidemia. The comparison of dyslipidemia conditions between the group with objective sleep duration ≥ 7 hours and the group with objective sleep duration < 7 hours showed a statistically significant difference （χ²=5.956， P<0.05）. Compared with the group with objective sleep duration ≥7 hours， the group with objective sleep duration < 7 hours exhibited significantly lower high-density lipoprotein cholesterol levels， and reduced sleep efficiency （t=-2.003， -5.482， P<0.05 or 0.01）. Binary Logistic regression analysis results showed that the risk of abnormal blood lipids in patients with chronic insomnia disorder with objective sleep duration < 7 hours was 3.128 times higher than that of patients with objective sleep duration ≥ 7 hours （OR=3.128， 95% CI： 1.139–8.588）. ConclusionObjective short sleep duration may be a risk factor for dyslipidemia in patients with chronic insomnia disorder.

BackgroundAnxiety symptoms have become a public health issue affecting the physical and mental health of the elderly population. Mental health literacy is a predictor of anxiety symptoms in the elderly. Currently， there are limited studies on the pathogenic mechanism between the two. Exploring the relationship and mechanism between mental health literacy and anxiety symptoms among the elderly is of great significance for improving the mental health level of the elderly. ObjectiveTo investigate the impact of mental health literacy on anxiety symptoms in the elderly， and to analyze the role of insomnia in this process， in order to provide references for the formulation of prevention and intervention strategies for anxiety symptoms in the elderly. MethodsFrom August 2021 to December 2022， a total of 10 650 older adults aged 60 years old and above were selected from a city in Sichuan Province using a multistage stratified sampling method. Participants completed the self-compiled demographic questionnaire， the Insomnia Severity Index （ISI）， the Generalized Anxiety Disorder Scale-7 item （GAD-7）， and the National Mental Health Literacy Questionnaire （NMHLQ）. Spearman correlation analysis was adopted to examine the correlation between the scores of the scales. Model 4 in SPSS 27.0 plugin Process 4.1 was employed to test the pathway of insomnia between mental health literacy （and its various dimensions） and anxiety symptoms. ResultsAmong the participants， 9 609 cases （90.23%） completed the valid questionnaire survey， and 1 680 cases （17.48%） were found to have anxiety symptoms. The total score of the NMHLQ for the elderly， as well as the scores of the knowledge， awareness， and skills dimensions， were negatively correlated with the GAD-7 score and the ISI score （r_s=-0.506–-0.054， P<0.01）， and the ISI score was positively correlated with the GAD-7 score （r_s=0.666， P<0.01）. Insomnia served as the mediating pathway between the mental health literacy and anxiety symptoms， with an indirect effect value of -0.210 （95% CI： -0.227–-0.193）， accounting for 54.97% of the total effect. Insomnia was the mediating pathway between the mental health literacy knowledge and anxiety symptoms of the elderly， with an indirect effect value of -0.161 （95% CI： -0.178–-0.144）， accounting for 52.61% of the total effect. Insomnia played a mediating role in the relationship between the awareness of mental health literacy and anxiety symptoms， with an indirect effect value of -0.323 （95% CI： -0.342–-0.302）， accounting for 76.36% of the total effect. Insomnia was the mediating pathway between the mental health literacy skills and anxiety symptoms of the elderly， with an indirect effect value of -0.172 （95% CI： -0.187–-0.159）， accounting for 53.75% of the total effect. ConclusionThe dimensions of mental health literacy， knowledge， awareness and skills of the elderly not only directly affect anxiety symptoms， but also indirectly influence anxiety symptoms through the pathway of insomnia.［Funded by Medical Research Project Plan of Sichuan Province （number， S23049）］

ObjectiveTo explore the diagnostic value of resting-state functional magnetic resonance imaging (rs-fMRI) for mild cognitive impairment (MCI) stage of Alzheimer's disease (AD), and to investigate the changes in brain functional connectivity in default mode network (DMN) and medial temporal lobe (MTL) regions. MethodsLiteratures were retrieved from multiple databases including CNKI, PubMed, Medline, and Embase, etc. from January, 2020 to October, 2025, investigating the changes in functional connectivity of DMN and MTL in patients with MCI compared to health control (HC) using rs-fMRI. Two researchers independently screened and extracted the literatures and methodological quality was assessed using QUADAS-2. Review Manager 5.4 was used to perform a meta-analysis of neuroimaging indicators in MCI patients and HC subjects. Stata and SDM were utilized to summarize diagnostic efficacy and brain functional connectivity, calculating the over all sensitivity, specificity and summary receiver operating characteristic-area under the curve (SROC-AUC). Deeks funnel plots were drawn, literature weights were analyzed, and subgroup analyses were conducted. ResultsA total of ten literatures were ultimately included, involving 1 010 patients with MCI and 1 714 HC subjects. MCI patients showed decreased or abnormal blood oxygen level dependence signals in DMN, focusing on bilateral medial prefrontal cortex (mPFC) and posterior cingulate cortex. The SROC-AUC was 0.91. In MTL, there was a characteristic decrease of spontaneous neural functional connectivities between the hippocampus and other regions, reflecting the obstruction of information transmission from episodic memory to the central nodes. Abnormal functional connectivity in DMN and MTL resulted in compensatory resting-state functional connectivity enhancement in other subnetworks or local functional connections, such as frontoparietal network and the hippocampal-parietal network. Abnormal activation of mPFC suggested atrophy of the hippocampus or abnormal brain function. The decline in functional connectivity between DMN and MTL indicated impairment of memory and information processing in the early stage. ConclusionThe early functional decoupling between DMN and MTL is a crucial neural mechanism at the MCI stage of AD, providing important neuroimaging evidence for the early diagnosis of AD.

ObjectiveTo quantitatively evaluate the effect of phrenic nerve-abdominal muscle electrical stimulation on pulmonary ventilation distribution in stroke patients using electrical impedance tomography (EIT). MethodsThirty-five stroke patients were enrolled at the First Affiliated Hospital of Chongqing Medical University from September, 2024 to June, 2025, and all received standardized phrenic nerve-abdominal muscle electrical stimulation. Percentage of ventilation in gravity-dependent regions of interesting (ROI%), center of ventilation (COV), global inhomogeneity index (GI) and change in end-expiratory lung impedance (ΔEELI) were measured with EIT monitoring before treatment (T0), immediately after treatment (T1), and at five, ten, 15, 20, 25, 30, 40 and 50 minutes after treatment (T2 to T9). ResultsThere were significant differences in ROI% (F = 7.003, P < 0.001) and COV (F = 5.722, P < 0.001) at different time points, both peaking at T1, followed by a downward trend until T5. No significant differences were observed in GI (F = 1.849, P = 0.097) and ΔEELI (F = 0.208, P = 0.871) across time points; however, GI at T7 was lower than that at T0 (P < 0.05). ConclusionPhrenic nerve-abdominal muscle electrical stimulation can improve the ventilation ratio in gravity-dependent regions and shift the center of ventilation dorsally. The improvement in ventilation distribution generally peaks at the end of treatment and lasts for approximately 20 minutes.

BackgroundPostpartum depression can affect the physical and mental health of mothers and the quality of parenting. Most Chinese women perform traditional postpartum practices （commonly known as "doing the month"） after giving birth， while the existing findings are inconsistent and inconclusive regarding the potential of traditional Chinese postpartum practices to alleviate or exacerbate postpartum depression. ObjectiveTo explore the relationship between traditional Chinese postpartum practices and postpartum depression， so as to provide references for reducing the risk of postpartum depression. MethodsA total of 240 consecutive women who gave birth in the obstetrics department of the Mianyang Central Hospital and the Third Hospital of Mianyang from January to May 2024 were selected. Data were collected using Self-designed General Information Questionnaire， Chinese version of the Edinburgh Postnatal Depression Scale （EPDS）， the Social Support Rating Scale （SSRS）， the Patient Health Questionnaire-15 （PHQ-15）， the Adherence to Doing-the-Month Practices questionnaire （ADP）， and the Self-compiled Questionnaire on the Cognition of Doing-the-Month. The absolute value （A value） of the difference between scores of ADP and Cognition of Doing-the-Month Questionnaire was calculated to evaluate the degree of cognitive behavioral conflict of postpartum women. Pearson correlation analysis was performed to examine the correlations of EPDS score with SSRS score， PHQ-15 score， ADP total and dimensional scores， Cognition of Doing-the-Month Questionnaire total and dimensional scores， and A value. Logistic regression analysis was conducted to identify the protective and risk factors for developing postpartum depression. ResultsThe postpartum depression was detected in 22.50% of women. The postpartum women had a EPDS score of （6.21±5.00）， ADP score of （70.05±20.57）， SSRS score of （41.96±6.96）， PHQ-15 score of （4.63±3.77）， and Cognition of Doing-the-Month questionnaire score of （40.30±10.13）. The A value was （0.65±0.58）. Correlation analysis revealed that EPDS score was negatively correlated with the total ADP score and the four dimensional scores of the restrictions on social activities， diet， housework， and personal hygiene （r=-0.228， -0.146， -0.184， -0.275， -0.168， P<0.05 or 0.01）， and positively correlated with the A value （r=0.161， P<0.05）. Logistic regression analysis indicated that restriction on housework dimension in ADP was entered into the model （OR=0.930， 95% CI： 0.885~0.978）. ConclusionThe restriction on housework dimension in traditional Chinese postpartum practices may be a protective factor against postpartum depression.

ObjectiveTo explore the evolution of medication patterns and syndrome-herb associations of traditional Chinese medicine （TCM） in treating inflammatory bowel disease （IBD）， providing a theoretical foundation for precise syndrome differentiation and treatment in clinical practice. MethodsMedical case literature on TCM treatment of IBD from 1960 to 2024 was retrieved to establish a database. Frequency statistics， cluster analysis， change point detection， and association rule mining were employed to comprehensively analyze the syndrome distribution， therapeutic methods， medication patterns， and their temporal variations. ResultsA total of 685 medical cases were included. Common syndromes were dampness-heat （66.42%） and spleen deficiency （56.20%）. Primary therapeutic methods included heat clearing （63.65%）， spleen invigorating （47.45%）， and dampness draining （36.79%）. High-frequency herbs included Coptidis Rhizoma （354）， Paeoniae Radix Alba （303）， Aucklandiae Radix （292）， Codonopsis Radix （253）， and Glycyrrhizae Radix et Rhizoma （244）. Initial prescription clustering revealed three core therapeutic method combinations： heat clearing and detoxifying （represented by Baitouweng Tang）， spleen invigorating and Qi reinforcing （represented by Shenling Baizhusan）， and cold-heat regulation （represented by Wumeiwan combined with Shaoyao tang）. Temporal analysis identified 2008 as a key transition point in TCM treatment of IBD， with significantly increased usage frequency of heat-clearing and dampness-drying herbs such as Fraxini Cortex， Phellodendri Chinensis Cortex， Sophorae Flavescentis Radix， and Scutellariae Radix as well as hemostatic herbs such as carbonized Sanguisorbae Radix， Bletillae Rhizoma， Agrimoniae Herba， and Notoginseng Radix et Rhizoma. Follow-up efficacy analysis showed median improvement rates of 64.0% at the first follow-up， 76.0% at the second follow-up， and 78.7% at the third follow-up. Syndrome-drug association analysis revealed specific herb pairs with significant therapeutic advantages， including Notoginseng Radix et Rhizoma + Coicis Semen， Sanguisorbae Radix + Coptidis Rhizoma， and Codonopsis Radix + Aconii Lateralis Radix Praeparaia. ConclusionTCM medication patterns for treating IBD demonstrate distinct temporal evolution characteristics， with significantly increased usage frequency of herbs such as Fraxini Cortex， Notoginseng Radix et Rhizoma， and Agrimoniae Herba. Significant therapeutic method-herb associations and syndrome-herb association patterns exist， with the formation of specific herb pairs， providing evidence-based support for precise syndrome differentiation and treatment of IBD.

Objective To obtain the data of radiological diagnosis and treatment resource distribution at medical institutions of different levels and in various cities, understand the status of resource allocation, provide policy-making basis and suggestions for optimizing the allocation of radiological diagnosis and treatment resources within the province, and offer data and references for related research. Methods A basic situation questionnaire survey was conducted on radiological diagnosis and treatment institutions in Hunan Province. Data were reviewed, analyzed, and statistically processed using Excel software to understand the allocation situation of radiological diagnosis and treatment resources in Hunan Province. Results As of 2022, there were 3031 radiological diagnosis and treatment institutions, 19770 radiation workers, and 6617 pieces of radiological diagnostic and treatment equipment. There were 80 radiation treatment institutions, 49 nuclear medicine institutions, 178 interventional radiology institutions, and 3028 X-ray imaging diagnosis institutions. There were 1077 radiation workers in radiation treatment, 461 radiation workers in nuclear medicine, 3369 radiation workers in interventional radiology, and 14,863 radiation workers in X-ray imaging diagnosis. There were 135 pieces of radiation treatment equipment, 56 pieces of nuclear medicine equipment, 262 pieces of interventional radiology equipment, and 6164 pieces of X-ray imaging diagnosis equipment. Conclusion Primary and lower grade medical institutions in Hunan Province account for a relatively high proportion of the total number of medical institutions, whereas radiological diagnostic and treatment resources are predominantly concentrated in tertiary medical institutions, demonstrating a unbalanced allocation of radiological diagnosis and treatment resources in various cities. In terms of the overall allocation of radiological diagnosis and treatment resources, interventional radiology is superior to the national average, nuclear medicine and X-ray imaging diagnosis are comparable to the national averages, and radiation treatment is lower than the national average. The total GDP is not the decisive factor in determining the development of local medical levels. It is suggested that relevant authorities should improve the rationality of the planning for the allocation of radiological diagnosis and treatment resources, promote the allocation of radiological diagnosis and treatment resources to the grassroots and underdeveloped areas through various measures, and thus achieve the goal of balanced resource allocation.